CN101595856A - A kind of freshwater rotifer common nutrient solution and method for making thereof - Google Patents

A kind of freshwater rotifer common nutrient solution and method for making thereof Download PDF

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Publication number
CN101595856A
CN101595856A CNA2009100122807A CN200910012280A CN101595856A CN 101595856 A CN101595856 A CN 101595856A CN A2009100122807 A CNA2009100122807 A CN A2009100122807A CN 200910012280 A CN200910012280 A CN 200910012280A CN 101595856 A CN101595856 A CN 101595856A
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China
Prior art keywords
phosphate buffer
nutrient solution
freshwater rotifer
solution
cobastab
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CNA2009100122807A
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Chinese (zh)
Inventor
殷旭旺
赵文
刘钢
魏杰
王丽
陈晓霞
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Dalian Fisheries University
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Dalian Fisheries University
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Priority to CNA2009100122807A priority Critical patent/CN101595856A/en
Publication of CN101595856A publication Critical patent/CN101595856A/en
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Abstract

The present invention relates to bio-culture solution.A kind of freshwater rotifer common nutrient solution adds following each raw material: potassium nitrate, bitter salt, six nitric hydrate calcium, dipotassium hydrogen phosphate, Cobastab in the phosphate buffer of pH7.0-7.5 12, biotin.The present invention joins with different proportionings and is mixed with the wheel animalcule culture fluid in the distilled water, is fit to cultivate common freshwater rotifer kind, and this culture fluid stability is high, and preparation is simple, and is nontoxic, and applicable scope is wider; For the indoor cultivation of the model organism wheel animalcule of experimental ecology research provides condition, also improved the training quality of indoor freshwater rotifer simultaneously, reduced the risk that wheel animalcule is cultivated.

Description

A kind of freshwater rotifer common nutrient solution and method for making thereof
Technical field
The present invention relates to bio-culture solution, particularly general indoor freshwater rotifer culture fluid.
Background technology
The freshwater rotifer of some kind as Brachionus calyciflorus (Brachionus calyciflorus), is internationally recognized model organism, can be indoor genecology, molecular biology and ecological toxicology research basic experiment material is provided.Because wheel animalcule is the live body biological subject, thereby need carry out long-term cultivation in the laboratory, experimentizes so that obtain the fresh and alive individuality of sufficient amount.The key technology of freshwater rotifer indoor cultivation is exactly the culture fluid of wheel animalcule.At present, most laboratory all is to cultivate wheel animalcule with the running water of aeration or the open-air pond water of filtration.But, more than two kinds of wheel animalcule culture fluids all have the problem of following several respects: (one) is because natural fresh water lacks buffer system, thereby the pH of culture fluid is extremely unstable, fluctuation on a large scale often occurs; (2) composition of the ion in the natural fresh water and ratio are usually unbalance, and some ionic species is on the high side, as Ca 2+, make that hardness, basicity or the chlorinity of culture fluid are higher; (3) environmental pollution often causes containing in the natural fresh water the higher toxicant of concentration (biotoxin, heavy metal and organic matter etc.), and these noxious materials are difficult to be removed by the method for aeration, filtration and high-temperature sterilization; (4) owing to the difference of environmental condition background value, its application of the natural fresh water in a certain area has limitation, can't be applicable to the freshwater rotifer that other areas collect, and does not have versatility.
Summary of the invention
The objective of the invention is the problem that exists in the above freshwater rotifer culture fluid in order to solve, a kind of freshwater rotifer common nutrient solution is provided, be fit to cultivate common freshwater rotifer kind, this culture fluid stability is high, nontoxic, and applicable scope is wider, improves the training quality of freshwater rotifer, reduces the risk that wheel animalcule is cultivated.
The technical scheme that the present invention is adopted for achieving the above object is: a kind of freshwater rotifer common nutrient solution adds following each raw material: potassium nitrate [KNO in the phosphate buffer of pH7.0-7.5 3], bitter salt [MgSO 47H 2O], six nitric hydrate calcium [Ca (NO 3) 26H 2O], dipotassium hydrogen phosphate [K 2HPO 4], Cobastab 12, biotin.
Add each raw material by following weight in the phosphate buffer of described pH 7.0-7.5: potassium nitrate 6-10g/l, bitter salt 3.7-6.2g/l, six nitric hydrate calcium 8.6-11.6g/l, dipotassium hydrogen phosphate 2.4-4.0g/l, Cobastab 120.5 μ g/l, biotin 0.5 μ g/l.
Described culture fluid preferably forms by following formulated:
The phosphate buffer of pH 7.3
KNO 3 8.0g/l
MgSO 4·7H 2O 4.96g/l
K 2HPO 4 3.2g/l
Ca(NO 3) 2·6H 2O 11.52g/l
Biotin 0.5 μ g/l
Cobastab 120.5 μ g/l.
Described culture fluid preferably forms by following formulated:
The phosphate buffer of pH 7.3
KNO 3 10.0g/l
MgSO 4·7H 2O 6.2g/l
K 2HPO 4 4.0g/l
Ca(NO 3) 2·6H 2O 14.4g/l
Biotin 0.5 μ g/l
Cobastab 120.5 μ g/l
Described culture fluid preferably forms by following formulated:
The phosphate buffer of pH 7.3
KNO 3 6.0g/l
MgSO 4·7H 2O 3.72g/l
K 2HPO 4 2.4g/l
Ca(NO 3) 2·6H 2O 8.64g/l
Biotin 0.5 μ g/l
Cobastab 120.5 μ g/l.
The method for making of described culture fluid: get NaH respectively 2PO 4Solution and Na 2HPO 4Solution is dissolved in the distilled water, is mixed with the phosphate buffer that meets the pH value; In phosphate buffer, add KNO then successively 3Solution, MgSO 47H 2O solution, K 2HPO 4Solution, Ca (NO 3) 26H 2O solution, and finally use the distilled water constant volume, in constant volume solution, add biotin and Cobastab at last respectively 12, promptly be mixed with freshwater rotifer common nutrient solution.
Freshwater rotifer culture fluid of the present invention is fit to cultivate common freshwater rotifer kind, this culture fluid prescription is reasonable, the stability of solution height, nontoxic, and applicable scope is wider, improve the training quality of freshwater rotifer, reduce the risk that wheel animalcule is cultivated, can be genecology, molecular biology and ecological toxicology model organism--wheel animalcule provides basic indoor cultivation environment.Culture fluid method for making of the present invention is simple.Through the wheel animalcule culture fluid that the advance copy invention is disposed, survival rate and the population growth rate of experiment wheel animalcule population all are significantly increased, and survival rate is the highest to improve 45%, and the population growth rate is the highest to improve 0.9.
Embodiment
Below in conjunction with specific embodiment the present invention is described in further detail, but the present invention is not limited to specific embodiment.
Embodiment 1
Prepare freshwater rotifer common nutrient solution as follows: get 2.3ml 0.2mol/LNaH at first respectively 2PO 4Solution and 7.7ml 0.2mol/L Na 2HPO 4Solution is dissolved in the distilled water of 500ml, is mixed with the phosphate buffer of pH value 7.3; The KNO that in this phosphate buffer, adds following each 8.0ml:10.0g/l of solution then successively 3The MgSO of solution---6.2g/l 47H 2The K of O solution---4.0g/l 2HPO 4Ca (the NO of solution---14.4g/l 3) 26H 2O solution, and finally use the distilled water constant volume to 1000ml, in this 1000ml solution, add biotin and the Cobastab of 0.5 μ g respectively 12, promptly be mixed with freshwater rotifer common nutrient solution.
Result of the test is as follows:
1. the wheel animalcule culture fluid that uses the present invention's configuration is after 10 days, the survival rate of Brachionus calyciflorus and population growth rate such as table 1.
Table 1
The survival rate of wheel animalcule (%) The population growth rate of wheel animalcule (individual/day)
The running water of aeration (control group) 51 0.41
The pond water (control group) that filters 69 0.63
Wheel animalcule culture fluid of the present invention 96 1.31
2. the wheel animalcule culture fluid that uses the present invention's configuration is after 15 days, the survival rate of ten index arm tail wheel worms and population growth rate such as table 2.
Table 2
The survival rate of wheel animalcule (%) The population growth rate of wheel animalcule (individual/day)
The running water of aeration (control group) 61 0.21
The pond water (control group) that filters 54 0.11
Wheel animalcule culture fluid of the present invention 89 0.81
3. the wheel animalcule culture fluid that uses the present invention's configuration is after 8 days, survival rate and population growth rate such as the table 3 of cornicult arm tail wheel worm.
Table 3
The survival rate of wheel animalcule (%) The population growth rate of wheel animalcule (individual/day)
The running water of aeration (control group) 15 0.03
The pond water (control group) that filters 30 0.09
Wheel animalcule culture fluid of the present invention 57 0.41
By above application example as can be seen, use the wheel animalcule culture fluid that the present invention disposed, survival rate and the population growth rate of experiment wheel animalcule population all are significantly increased, and survival rate is the highest to improve 45%, and the population growth rate is the highest to improve 0.9.
Embodiment 2
Prepare freshwater rotifer common nutrient solution according to embodiment 1 described method, at NaH 2PO 4And Na 2HPO 4Add following raw material in the phosphate buffer of the pH 7.3 of preparation, each material quantity is as follows in the finished product:
KNO 3 10.0g/l
MgSO 4·7H 2O 6.2g/l
K 2HPO 4 4.0g/l
Ca(NO 3) 2·6H 2O 14.4g/l
Biotin 0.5 μ g/l
Cobastab 120.5 μ g/l.
Embodiment 3
Prepare freshwater rotifer common nutrient solution according to embodiment 1 described method, at NaH 2PO 4And Na 2HPO 4Add following raw material in the phosphate buffer of the pH 7.3 of preparation, each material quantity is as follows in the finished product:
KNO 3 6.0g/l
MgSO 4·7H 2O 3.72g/l
K 2HPO 4 2.4g/l
Ca(NO 3) 2·6H 2O 8.64g/l
Biotin 0.5 μ g/l
Cobastab 120.5 μ g/l.
Embodiment 4
Prepare freshwater rotifer common nutrient solution according to embodiment 1 described method, at NaH 2PO 4And Na 2HPO 4The raw material that adds embodiment 1 described consumption in the phosphate buffer of the pH 7.0 of preparation.
Embodiment 5
Prepare freshwater rotifer common nutrient solution according to embodiment 1 described method, at NaH 2PO 4And Na 2HPO 4The raw material that adds embodiment 2 described consumptions in the phosphate buffer of the pH 7.0 of preparation.
Embodiment 6
Prepare freshwater rotifer common nutrient solution according to embodiment 1 described method, at NaH 2PO 4And Na 2HPO 4The raw material that adds embodiment 3 described consumptions in the phosphate buffer of the pH 7.0 of preparation.
Embodiment 7
Prepare freshwater rotifer common nutrient solution according to embodiment 1 described method, at NaH 2PO 4And Na 2HPO 4The raw material that adds embodiment 1 described consumption in the phosphate buffer of the pH 7.5 of preparation.
Embodiment 8
Prepare freshwater rotifer common nutrient solution according to embodiment 1 described method, at NaH 2PO 4And Na 2HPO 4The raw material that adds embodiment 2 described consumptions in the phosphate buffer of the pH 7.5 of preparation.
Embodiment 9
Prepare freshwater rotifer common nutrient solution according to embodiment 1 described method, at NaH 2PO 4And Na 2HPO 4The raw material that adds embodiment 3 described consumptions in the phosphate buffer of the pH 7.5 of preparation.

Claims (6)

1, a kind of freshwater rotifer common nutrient solution is characterized in that: add following each raw material in the phosphate buffer of pH7.0-7.5: potassium nitrate, bitter salt, six nitric hydrate calcium, dipotassium hydrogen phosphate, Cobastab 12, biotin.
2, a kind of freshwater rotifer common nutrient solution according to claim 1 is characterized in that: add each raw material by following weight in the phosphate buffer of pH7.0-7.5: potassium nitrate 6-10g/l, bitter salt 3.7-6.2g/l, six nitric hydrate calcium 8.6-11.6g/l, dipotassium hydrogen phosphate 2.4-4.0g/l, Cobastab 120.5 μ g/l, biotin 0.5 μ g/l.
3, a kind of freshwater rotifer common nutrient solution according to claim 1 is characterized in that: the raw material that contains following weight in the phosphate buffer of pH 7.3:
KNO 3 8.0g/l
MgSO 4·7H 2O 4.96g/l
K 2HPO 4 3.2g/l
Ca(NO 3) 2·6H 2O 11.52g/l
Biotin 0.5 μ g/l
Cobastab 120.5 μ g/l.
4, a kind of freshwater rotifer common nutrient solution according to claim 1 is characterized in that: the raw material that contains following weight in the phosphate buffer of pH 7.3:
The phosphate buffer of pH 7.3
KNO 3 10.0g/l
MgSO 4·7H 2O 6.2g/l
K 2HPO 4 4.0g/l
Ca(NO 3) 2·6H 2O 14.4g/l
Biotin 0.5 μ g/l
Cobastab 120.5 μ g/l.
5, a kind of freshwater rotifer common nutrient solution according to claim 1 is characterized in that: the raw material that contains following weight in the phosphate buffer of pH 7.3:
The phosphate buffer of pH 7.3
KNO 3 6.0g/l
MgSO 4·7H 2O 3.72g/l
K 2HPO 4 2.4g/l
Ca(NO 3) 2·6H 2O 8.64g/l
Biotin 0.5 μ g/l
Cobastab 120.5 μ g/l.
6, the method for making of the arbitrary described a kind of freshwater rotifer common nutrient solution of claim 1-5 is characterized in that: get NaH respectively 2PO 4Solution and Na 2HPO 4Solution is dissolved in the distilled water, is mixed with the phosphate buffer that meets the pH value; In phosphate buffer, add KNO then successively 3Solution, MgSO 47H 2O solution, K 2HPO 4Solution, Ca (NO 3) 26H 2O solution, and finally use the distilled water constant volume, in constant volume solution, add biotin and Cobastab at last respectively 12, promptly be mixed with freshwater rotifer common nutrient solution.
CNA2009100122807A 2009-06-25 2009-06-25 A kind of freshwater rotifer common nutrient solution and method for making thereof Pending CN101595856A (en)

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Cited By (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN102246722A (en) * 2011-06-10 2011-11-23 浙江海洋学院 High density rotifer culture device
CN102599109A (en) * 2012-02-29 2012-07-25 谢达祥 Simple method for purifying rotifers in batch
CN103141441A (en) * 2012-10-23 2013-06-12 中国水产科学研究院东海水产研究所 Method for cultivating rotifers by using dried yeast and Antarctic krill meal
CN104082248A (en) * 2014-06-23 2014-10-08 浙江省淡水水产研究所 Method for small-scale and simple culturing of freshwater rotifers

Cited By (7)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN102246722A (en) * 2011-06-10 2011-11-23 浙江海洋学院 High density rotifer culture device
CN102246722B (en) * 2011-06-10 2013-03-06 浙江海洋学院 High density rotifer culture device
CN102599109A (en) * 2012-02-29 2012-07-25 谢达祥 Simple method for purifying rotifers in batch
CN103141441A (en) * 2012-10-23 2013-06-12 中国水产科学研究院东海水产研究所 Method for cultivating rotifers by using dried yeast and Antarctic krill meal
CN103141441B (en) * 2012-10-23 2014-06-04 中国水产科学研究院东海水产研究所 Method for cultivating rotifers by using dried yeast and Antarctic krill meal
CN104082248A (en) * 2014-06-23 2014-10-08 浙江省淡水水产研究所 Method for small-scale and simple culturing of freshwater rotifers
CN104082248B (en) * 2014-06-23 2016-01-20 浙江省淡水水产研究所 The method of the simple and easy cultivation freshwater rotifer of a kind of small-scale

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Application publication date: 20091209